RESEARCH ARTICLE
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Molecular responses and chromosomal aberrations in patients with polycythemia vera treated with peg-proline-interferon alpha-2b Nicole C.C. Them,1 Klaudia Bagienski,1 Tiina Berg,1 Bettina Gisslinger,2 Martin Schalling,2 Doris Chen,1 Veronika Buxhofer-Ausch,3,4 Josef Thaler,5 Ernst Schloegl,6 Guenther A. Gastl,7 Dominik Wolf,7,8 Karin Strecker,3 Alexander Egle,9 Thomas Melchardt,9 Sonja Burgstaller,5 Ella Willenbacher,7 Oleh Zagrijtschuk,10 Christoph Klade,10 Richard Greil,9 Heinz Gisslinger,2 and Robert Kralovics1,2* Fifty-one polycythemia vera (PV) patients were enrolled in the phase I/II clinical study PEGINVERA to receive a new formulation of pegylated interferon alpha (peg-proline-IFNa-2b, AOP2014/P1101). Peg-proline-IFNa-2b treatment led to high response rates on both hematologic and molecular levels. Hematologic and molecular responses were achieved for 46 and 18 patients (90 and 35% of the whole cohort), respectively. Although interferon alpha (IFNa) is known to be an effective antineoplastic therapy for a long time, it is currently not well understood which genetic alterations influence therapeutic outcomes. Apart from somatic changes in specific genes, large chromosomal aberrations could impact responses to IFNa. Therefore, we evaluated the interplay of cytogenetic changes and IFNa responses in the PEGINVERA cohort. We performed high-resolution SNP microarrays to analyze chromosomal aberrations prior and during peg-proline-IFNa-2b therapy. Similar numbers and types of chromosomal aberrations in responding and non-responding patients were observed, suggesting that peg-proline-IFNa-2b responses are achieved independently of chromosomal aberrations. Furthermore, complete cytogenetic remissions were accomplished in three patients, of which two showed more than one chromosomal aberration. These results imply that peg-proline-IFNa-2b therapy is an effective drug for PV patients, possibly including patients with complex cytogenetic changes. C 2014 The Authors. American Journal of Hematology published by Wiley Periodicals, Inc. Am. J. Hematol. 90:288–294, 2015. V
䊏 Introduction Polycythemia vera (PV) represents a frequent type of BCR-ABL negative myeloproliferative neoplasm (MPN) and is characterized by an elevated erythrocyte mass, as well as a variable presence of thrombocytosis and leukocytosis [1,2]. PV patients have a risk of developing thrombosis, bleeding and disease transformation to secondary myelofibrosis as well as secondary acute myeloid leukemia. In the vast majority (95%) of PV patients the JAK2-V617F mutation is found and in around 3% of PV patients the less common JAK2 exon 12 mutations [3–9]. The functional consequences of these mutations are a constitutively active JAK2 kinase signaling and cytokine hypersensitivity of myeloid cells [5,8,10–12]. Present treatment options for PV patients include phlebotomy, low-dose aspirin, hydroxyurea and interferon alpha (IFNa) [2]. Already in the 1980s therapeutic efficacy of recombinant IFNa for MPN patients was reported [13–15]. IFNa not only induced hematologic and molecular responses in most MPN patients but was also described to be nonleukemogenic [16]. However, the use of recombinant IFNa was limited by frequent drugrelated toxicities and subsequent treatment discontinuations. The chemical linkage of polyethylene glycol (peg) to IFNa increased the plasma half-life and improved the toxicity profile of peg-IFNa considerably [17]. To date, the cellular and molecular mechanisms involved in the myelosuppressive
Additional Supporting Information may be found in the online version of this article. The copyright line for this article was changed on 20 August 2015 after original online publication. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. 1
CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria; 2Department of Internal Medicine I, Division of Hematology and Blood Coagulation, Medical University of Vienna, Vienna, Austria; 32nd Medical Department, Sozialmedizinisches Zentrum Ost—Donauspital, Vienna, Austria; 4 Interne 1 Hemato-Oncology, Krankenhaus Der Elisabethinen Linz, Linz, Austria; 5Department of Internal Medicine IV, Wels-Grieskirchen Hospital, Wels, Austria; 6 Third Medical Department, Hanusch Hospital, Vienna, Austria; 7Department of Internal Medicine V, Haematology & Oncology, Innsbruck Medical University, Innsbruck, Austria; 8Medical Clinic III, Oncology, Hematology and Rheumatology, University Clinic of Bonn (UKB), Bonn, Germany; 9Laboratory for Immunological and Molecular Cancer Research, Department of Internal Medicine III with Hematology, Medical Oncology, Hemostaseology, Infectious Diseases, Rheumatology, Oncologic Center, Paracelsus Medical University, Salzburg, Austria; 10AOP Orphan Pharmaceuticals AG, Vienna, Austria
Conflict of interest: All authors, except Zagrijtschuk and Klade, received research funding from AOP Orphan Pharmaceuticals AG. Zagrijtschuk and Klade are employed by AOP Orphan Pharmaceuticals AG. *Correspondence to: Robert Kralovics, PhD, CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT25.3, 1090 Vienna, Austria. E-mail:
[email protected] Contract grant sponsor: Austrian Science Fund; Contract grant numbers: FWF4702-B20; FWF20812-B20. Contract grant sponsor: AOP Orphan Pharmaceuticals AG. Received for publication: 18 December 2014; Accepted: 20 December 2014 Am. J. Hematol. 90:288–294, 2015. Published online: 24 December 2014 in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/ajh.23928 C 2014 The Authors. American Journal of Hematology published by Wiley Periodicals, Inc. V
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American Journal of Hematology, Vol. 90, No. 4, April 2015
doi:10.1002/ajh.23928
RESEARCH ARTICLE effect of IFNa remain as unclear as the influence of constitutive and acquired genetic changes on the magnitude of IFNa responses. To evaluate the impact of acquired chromosomal aberrations we analyzed chromosomal lesions by high-resolution SNP microarrays in PV patients treated with a new formulation of peg-IFNa (AOP2014/P1101, peg-proline-IFNa-2b) in a phase I/II clinical trial.
䊏 Methods Patient samples. Patients were enrolled in the phase I/II clinical study PEGINVERA (identifier: NCT01193699; www.clinicaltrials.gov) [18,19]. The study was approved by the Institutional Review Board of the Medical University of Vienna and the Austrian Drug Agency (AGES) and all patients provided written consent. Granulocytes were isolated from peripheral blood by standard gradient centrifugation using Histopaque 1077 (Sigma–Aldrich, St. Louis, MO). Genomic DNA was isolated from granulocytes and whole blood samples using the Wizard Genomic DNA Purification Kit and the Maxwell 16 Tissue DNA Purification Kit (Promega, Madison, WI). Hematologic response criteria. Hematologic response was defined similar to the European LeukemiaNet (ELN) criteria [20]. Complete hematologic response (CHR) required hematocrit
